QTL mapping of carrot resistance to leaf blight with connected populations: stability across years and consequences for breeding

Combining biparental and multiparental connected population analyses was useful for the identification of 11 QTLs in two new genetic backgrounds of carrot resistance to Alternaria dauci and for breeding recommendations. Leaf blight due to the fungus Alternaria dauci is the major carrot foliar disease worldwide. Some resistance QTLs have been previously identified in one population, but the evaluation of additional genetic backgrounds with higher level of resistance would give opportunities for breeders to combine them by pyramiding. For this purpose, two segregating populations were evaluated twice across 4 years in the same environment (1) to compare the efficiency of the single vs. the connected populations approach for characterizing the new sources of carrot resistance to Alternaria dauci; (2) to evaluate the stability of QTLs over the years; and (3) to give recommendations to breeders for marker-assisted selection. Single and connected analyses were complementary; their combination allowed the detection of 11 QTLs. Connected analyses allowed the identification of common and specific QTLs among the two populations and the most favorable allele at each QTL. Important contrasts between allelic effects were observed with four and five most favorable alleles coming from the two resistant parental lines, whereas two other favorable alleles came from the susceptible parental line. While four QTLs were consistent across years, seven were detected within a single year. The heritabilities for both populations PC2 and PC3 were high (75 and 78 %, respectively), suggesting that the resistance of carrot to A. dauci was little affected by these environmental conditions, but the instability of QTL over years may be due to changing environmental conditions. The complementarity between these parental lines in terms of interesting allelic combinations is also discussed

Improved liquid chromatographic method for determination of carotenoids in carrot

Carotenoids are a large class of plant metabolites with a function of either essential nutrients or health promoting compounds for humans. Carrot root is a well-known and significant source of dietary carotenoids, mainly: α- and β-carotene, lutein and lycopene. These pigments are the main carotenoids separated and quantified routinely by HPLC analysis. However, little is known about minor carotenoids, carotenoid esters and the carotenoids present in leaves despite their potential interest in metabolic and physiological studies. Previous works used C-18 columns but these stationary phases provide a poor resolution of structurally similar compounds and geometrical isomers. In recent years, C-30 columns have been developed and successfully applied at the separation of carotenoids from various plant materials, the number of resolved carotenoids being significantly improved. Based on literature procedures, we have developed a HPLC-DAD method with a C-30 column, adapted to the quantification of carotenoid compounds from carrot roots and leaves. A simple and rapid extraction method was optimized for both these types of samples on a panel of 5 genotypes displaying distinct root colours (different carotenoid composition and contents). Carotenoids from roots were separated in 23 minutes while carotenoids and chlorophylls from leaves were separated in 42 minutes. Compounds were identified according to their retention time and UV-visible spectrum in comparison with authentic standards (analysed individually and in combination, in the same conditions), or with data from literature, when standards were unavailable. Results showed that carrot root exhibited a simple profile with only 1 to 3 main carotenoids whereas a more complex composition was noticed in leaves, containing both identified and unidentified carotenoids and chlorophylls. Moreover, the composition was quite conservative for leaves but depended on the genotype for roots

Effect of GA 3 and paclobutrazol on adventitious shoot regeneration of two Pelargonium sp

This study had two aims. The first was to improve the regeneration efficiency of Pelargonium leaf discs by adventitious budding. The second was to test the effect of gibberellic acid (GA 3) and paclobutrazol (PBZ) on callus formation and adventitious shoot regeneration in Pelargonium before using genetic transformation of this species for functional validation of genes involved in the process of GA regulation. GA 3 and paclobutrazol (an inhibitor of GA synthesis pathway) were added (together or separately) in the shoot regeneration media of two Pelargonium species, Pelargonium * hortorum \u27Panache sud\u27 (\u27P.sud\u27) and Pelargonium * domesticum \u27Autumn haze\u27 (\u27 P.dom\u27). In both cases, GA 3 applied alone, completely inhibited the bud regeneration. Moreover, the rate of callus formation decreased drastically when 5 M of GA 3 was applied to \u27 P. dom\u27 explants. Similar result was obtained with \u27P.sud\u27 explants using 20 M GA 3. Paclobutrazol (0.3 M) applied at the same time as GA 3 (10 M) could partially restore regeneration process of \u27 P. dom\u27. For \u27 P. dom\u27, the use of paclobutrazol alone increased callus formation and slightly improved the rate of regeneration. Moreover, initiated buds had a better appearance. For \u27P. sud\u27, which had an abundant callusing, paclobutrazol did not improve regeneration and led to hyperhydric shoots

Carotenoid gene expression explains the difference of carotenoid accumulation in carrot root tissues

The carrot root is well divided into two different tissues separated by vascular cambium: the secondary phloem and xylem. The equilibrium between these two tissues represents an important issue for carrot quality, but the knowledge about the respective carotenoid accumulation is sparse. The aim of this work was (i) to investigate if variation in carotenoid biosynthesis gene expression could explain differences in carotenoid content in phloem and xylem tissues and (ii) to investigate if this regulation is differentially modulated in the respective tissues by water-restricted growing conditions. In this work, five carrot genotypes contrasting by their root color were studied in control and water-restricted conditions. Carotenoid content and the relative expression of 13 genes along the carotenoid biosynthesis pathway were measured in the respective tissues. Results showed that in orange genotypes and the purple one, carotenoid content was higher in phloem compared to xylem. For the red one, no differences were observed. Moreover, in control condition, variations in gene expression explained the different carotenoid accumulations in both tissues, while in water-restricted condition, no clear association between gene expression pattern and variations in carotenoid content could be detected except in orange-rooted genotypes. This work shows that the structural aspect of carrot root is more important for carotenoid accumulation in relation with gene expression levels than the consequences of expression changes upon water restriction

Interplay of Sugar, Light and Gibberellins in Expression of Rosa hybrida Vacuolar Invertase 1 Regulation

Our previous findings showed that the expression of the Rosa hybrida vacuolar invertase 1 gene (RhVI1) was tightly correlated with the ability of buds to grow out and was under sugar, gibberellin and light control. Here, we aimed to provide an insight into the mechanistic basis of this regulation. In situ hybridization showed that RhVI1 expression was localized in epidermal cells of young leaves of bursting buds. We then isolated a 895 bp fragment of the promoter of RhVI1. In silico analysis identified putative cis-elements involved in the response to sugars, light and gibberellins on its proximal part (595 bp). To carry out functional analysis of the RhVI1 promoter in a homologous system, we developed a direct method for stable transformation of rose cells. 5′ deletions of the proximal promoter fused to the uidA reporter gene were inserted into the rose cell genome to study the cell’s response to exogenous and endogenous stimuli. Deletion analysis revealed that the 468 bp promoter fragment is sufficient to trigger reporter gene activity in response to light, sugars and gibberellins. This region confers sucrose- and fructose-, but not glucose-, responsive activation in the dark. Inversely, the –595 to –468 bp region that carries the sugar-repressive element (SRE) is required to down-regulate the RhVI1 promoter in response to sucrose and fructose in the dark. We also demonstrate that sugar/light and gibberellin/light act synergistically to up-regulate β-glucuronidase (GUS) activity sharply under the control of the 595 bp pRhVI1 region. These results reveal that the 127 bp promoter fragment located between –595 and –468 bp is critical for light and sugar and light and gibberellins to act synergistically

Shoot regeneration and genetic transformation by Agrobacterium tumefaciens of Hydrangea macrophylla Ser. leaf discs

A reproducible procedure was developed for genetic transformation of Hydrangea macrophylla Ser. cv. Blaumeise by Agrobacterium tumefaciens following the development of an efficient regeneration system using leaf discs excised from 12 to 15 weeks old meristem-derived vitroplants. Explants were cultivated on solid B5 medium complemented with maltose 110 mM, BAP 10 μM and NAA 0.5 μM. A low light regime of 17 μmol m−2 s−1 improved regeneration frequency up to 86%. For transformation, leaf discs were inoculated and co-cultivated with two disarmed A. tumefaciens strains, EHA 101 and LBA 4404, both carrying the binary vector pFAJ3000 which contained the nptII selectable gene and the GUS reporter gene. A pre-culture period of 3 days and a short co-cultivation duration (1 day) improved the efficiency of transformation. Inoculation of only 10 min with agitation including (or not) vacuum infiltration was sufficient. If selection on kanamycin containing medium was applied after a 2 weeks culture period on shoot regeneration medium, the percentage of explants forming kanamycin-resistant shoots increased from 3.3 to 13.3%. Integration and expression of the introduced transgene were confirmed by histochemical GUS assay, PCR and Southern blot analysis. Flowering of transgenic plants in glasshouse occurred 10 months after acclimatization

Omega 3, 6, 9 Enhanced Goat Meat (Omega-Chevon) from Flaxseed and Canola Fed Meat Goats

With growing obesity and cardiovascular disease concerns, the meat industry aims to reduce fat content in meat products. Currently Omega Fatty Acid (FA) enhanced beef and eggs are being marketed in the US, but Omega enhanced goat meat (Omega-Chevon) has not been developed. Meat goats were fed ground flaxseed and canola supplemented feed for 90 days. There were no palatability, weight, or health issues in meat goats fed canola and flaxseed supplemented feed. Chevon from goats fed canola and flaxseed had significant (

Development of Cryopreservation Protocols for ex-vitro-grown Rosa Shoot-tips (Rosa chinensis ‘Old Blush’)

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